Mutational analysis of the J recombination signal sequence binding protein (RBP-J)/Epstein-Barr virus nuclear antigen 2 (EBNA2) and RBP-J/Notch interaction
Mutational analysis of the J recombination signal sequence binding protein (RBP-J)/Epstein-Barr virus nuclear antigen 2 (EBNA2) and RBP-J/Notch interaction
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DOI：
10.1046/j.1432-1327.2001.02387.x
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发表时间：
2001-09-01
期刊：
影响因子：
--
通讯作者：
Kempkes, B
中科院分区：
文献类型：
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作者：
Fuchs, KP;Bommer, G;Kempkes, B
EB病毒核抗原2（EBNA 2）和Notch蛋白都在细胞核内作为转录衔接蛋白发挥作用。EBNA 2在EB病毒（EBV）对原代B细胞的永生化过程中起关键作用。Notch蛋白参与淋巴瘤发生以及组织分化和发育过程中的多细胞命运决定。EBNA 2和Notch两者都与DNA结合蛋白RBP-J相互作用，从而接近其靶基因的启动子。为了鉴定J重组信号序列结合蛋白（RBP-J）中与Notch或EBNA 2相互作用相关的区域，我们对RBP-J进行了突变分析。通过反向双杂交系统筛选RBP-J突变体文库中不能与EBNA 2或Notch结合的等位基因。这些等位基因的序列分析表明，一个有限的和特别独特的数量的氨基酸位置是相关的相互作用。鉴于RBP-J在B细胞永生化中的重要作用，EBNA 2/RBP-J蛋白-蛋白相互作用可能是EBV相关疾病治疗干预的候选靶标。
Epstein-Barr virus nuclear antigen 2 (EBNA2) and the Notch protein both function within the nucleus as transcriptional adaptor proteins. EBNA2 plays a key role during the immortalization of primary B-cells by Epstein-Barr virus (EBV). Notch proteins are involved in lymphomagenesis as well as in multiple cell fate decisions during tissue differentiation and development. Both, EBNA2 and Notch interact with the DNA binding protein RBP-J and thereby gain access to the promoter of their target genes. In order to identify regions within the J recombination signal sequence binding protein (RBP-J), that are relevant for either the Notch or the EBNA2 interaction, we have performed a mutational analysis of RBP-J. A library of RBP-J mutants was screened by a reverse two-hybrid system for alleles that fail to bind to either EBNA2 or Notch. The sequence analysis of these alleles reveals that a limited and particularly distinct number of amino-acid positions are relevant for either interaction only. Given the important role of RBP-J in B-cell immortalization, the EBNA2/RBP-J protein-protein interaction could be a candidate target for therapeutic intervention in EBV related diseases.